• Journal of Life Science
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• v.28 no.5
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• pp.532-539
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• 2018

Hair graying is processed by loss of melanin production caused by the decrease of activity and number of melanocyte and the accumulation of hydrogen peroxide ($H_2O_2$) in the hair follicle with increase of age. The purpose of this study was to investigate the effect the Black oryzasativa ethanolic extract (BLEE) on the melanin production. In this study BLEE at $8{\mu}g/ml$ or more showed a significant 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity and reduction power. BLEE at $16{\mu}g/ml$ or more showed promoted tyrosinase activity and melanin production. In addition BLEE scavenged intracellular $H_2O_2$ in 2',7'-dichlorodihydrofluorescein (DCF) fluorescence assay in B16F1 cells. However, Western blot analyses displayed that BLEE decreased the expression level of catalase, but no effect on the expression level of tyrosinase, tyrosinase associated protein-1 (TRP-1), tyrosinase associated protein-2 (TRP-2) and microphthalmia-associated transcription factor (MITF) transcription factor involved in melanogenesis. Thus, the promotive effect of BLEE on melanin production is attributed to the increase of tyrosinase activity and the reduction of intracellular $H_2O_2$ level. In conclusion, BLEE played a key role in in promoting melanin production, which suggests that the BLEE could be applied as a potential functional material in the development of hair care cosmetics related to the promotion of melanin production for the growth of black hair.

• Journal of Microbiology and Biotechnology
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• v.28 no.3
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• pp.375-380
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• 2018

We have previously found that mycelia culture broth of eight kinds of traditional herbal extracts fermented with Phellinus linteus (previously named as 8-HsPLCB) not only inhibited melanin and tyrosinase activity, but also reduced the contents of melanogenesis-related proteins, including tyrosinase and microphthalmia-associated transcription factor, in 3-isobutyl-1-methylxanthine-stimulated B16F0 melanoma cells. For a further study, the effect of 8-HsPLCB against skin pigmentation in brown guinea pigs with ultraviolet B (UVB)-induced hyperpigmentation was investigated. 8-HsPLCB (3%) and arbutin (2%) as positive controls were applied topically twice daily for 4 weeks to the hyperpigmented areas. 8-HsPLCB showed skin-lightening effect as effective as arbutin, one of the most widely used in whitening cosmetics. Melanin index values as the degree of pigmentation showed a significant reduction week by week post 8-HsPLCB treatment and then substantially reduced by 4 weeks. The degree of depigmentation after 4 weeks of topical application with 8-HsPLCB was 32.2% as compared with before treatment (0 week). Moreover, using Fontana-Masson staining and hematoxylin-eosin staining, 8-HsPLCB reduced melanin pigmentation in the basal layer of the epidermis and epidermal thickness changes exposed to the UV-B irradiation as compared with non-treatment and vehicle treatment. The intensity of the skin-lightening effect of 8-HsPLCB was similar to arbutin. These results suggest that the skin-lightening effect of 8-HsPLCB might be resulted from inhibition of melanin synthesis by tyrosinase in melanocytes. To conclude, 8-HsPLCB treatment showed reduction of the melanin pigment and histological changes induced by UV irradiation in brown guinea pigs.

• Journal of Life Science
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• v.27 no.12
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• pp.1452-1461
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• 2017

Melanin production by melanocytes in human hair follicles decreases with time and leads to the graying process, which is a phenotype of human aging and an index of aging. The reduction in melanin production is the result of decreased tyrosinase activity in hair follicles and an accumulation of active oxygen species, such as hydrogen peroxide. This study investigated antioxidant effects and melanin-promoting effects in B16F1 cells treated with black sesame ethanolic nonpolar-soluble extract (SBEEO) and black sesame ethanolic polar-soluble extract (SBEEP). In antioxidation experiments, both SBEEP and SBEEO did not eliminate 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical, but SBEEO at $64{\mu}g/ml$ showed low reducing power. SBEEP exerted cytotoxic effects at concentrations greater than $8{\mu}g/ml$, whereas SBEEO showed cytotoxic effects at concentrations greater than $4{\mu}g/ml$. SBEEP and SBEEO induced melanin synthesis, tyrosinase activity, and DOPA oxidase activity in vitro. In live cells, melanin synthesis was greater in the SBEEP treatment group as compared with that in the SBEEO treatment group. SBEEP stimulated melanin synthesis by modulating the expression of tyrosinase-related protein-2 (TRP-2), which is an important enzyme in melanin synthesis. These results imply that SBEEP obtained from black sesame ethanolic extract may have the potential to improve melanin synthesis.

• Journal of Microbiology and Biotechnology
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• v.30 no.8
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• pp.1184-1194
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• 2020

Melanin is a major factor that darkens skin color as one of the defense systems to prevent the harmful effects of UV light. However, darkened skin from the localized or systemic accumulation of melanin is viewed in many cultures as an esthetic problem. Consequentially, searching for anti-melanogenic agents from natural sources is very popular worldwide. Previous screening of fermented rice products, obtained from various rice cultivars fermented with different sources of loog-pang (Thai traditional fermentation starter), revealed that the highest ability to reduce the melanin content in B16F10 melanoma cells was from unpolished black rice fermented with a defined starter mixture of microbes isolated from loog-pang E11. The aim of this study was to investigate the mechanism of the fermented unpolished black rice (FUBR) on the inhibition of melanogenesis in B16F10 melanoma cells. The strongest reduction of cellular melanin content was found in the FUBR sap (FUBRS). The melanin reduction activity was consistent with the significant decrease in the intracellular tyrosinase activity. The FUBRS showed no cytotoxic effect to B16F10 melanoma or Hs68 human fibroblast cell lines. It also significantly reduced the transcript and protein expression levels of tyrosinase, tyrosinase-related protein 1 (TYRP-1), TYRP-2, and microphthalmia-associated transcription factor. Furthermore, it induced a significantly increased level of phosphorylated ERK, p38 and Akt signaling pathways, which likely contributed to the negative regulation of melanogenesis. From these results, a model for the mechanism of FUBRS on melanogenesis inhibition was proposed. Moreover, these results strongly suggested that FUBRS possesses anti-melanogenesis activity with high potential for cosmeceutical application as a skin depigmenting agent.

• YAKHAK HOEJI
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• v.57 no.1
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• pp.24-31
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• 2013

Although statins, inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, have been shown to increase melanin synthesis, the exact mechanism of this action is not fully understood. In this study we investigated the possible involvement of intracellular $Ca^{2+}$ signal in the mechanism of stimulation of melanin synthesis induced by lovastatin in B16 cells. Lovastatin stimulated the production of melanin in a dose-dependent manner in the cells. Treatment with mevalonate, FPP and GGPP, precursors of cholesterol, did not significantly suppress the lovastatin-induced melanin production, suggesting that inhibition of cholesterol synthesis may not be involved in the mechanism of the action of lovastatin. In addition, lovastatin did not significantly alter the cAMP concentration and the stimulated production of melanin by lovastatin was not significantly changed by treatment with H89, a potent inhibitor of protein kinase A, which demonstrates that cAMP pathway may not be involved. However, lovastatin increased intracellular $Ca^{2+}$ concentration in a dose-related fashion. Treatment with EGTA, an extracellular $Ca^{2+}$ chelator did not significantly alter the lovastatin-induced intracellular $Ca^{2+}$ increase and melanin synthesis, whereas intracellular $Ca^{2+}$ reduction with BAPTA/AM and intracellular $Ca^{2+}$ release blockers (dantrolene and TMB-8) completely blunted these actions of lovastatin. Taken together, these results suggest that the intracellular $Ca^{2+}$ release may play an important role in the lovastatin-induced stimulation of melanin synthesis in B16 cells. These results further suggest that lovastatin may be useful for the treatment of hypopigmentation disorders, such as vitiligo.

• Journal of Life Science
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• v.34 no.4
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• pp.236-244
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• 2024

Melanin is a natural pigment found in most plants and animals, and it is involved in determining the color of the skin and hair. Melanogenesis is a reactive occurrence in melanocytes aiming to protect the skin from external stimuli, such as ultraviolet rays. Tyrosine is involved in the biosynthesis of the substrate tyrosine into melanin. However, melanin overproduction can lead to skin diseases, such as melasma, blotching, hyperpigmentation, and skin cancer. Although many studies have been conducted on whitening substances, such as kojic acid and arbutin, some countries have banned or refrained from using them due to their side effects. Therefore, this study assessed the potential of horseradish (HR) as a new whitening agent in cosmetic products. For efficient extraction, subcritical water extraction was conducted. The results showed that the horseradish subcritical water 200℃ (SW 200) extract showed high DPPH radical scavenging ability, total phenolic contents (TPC), inhibiting tyrosinase activity and inhibiting melanin production of B16-F10 melanoma cell lines. To investigate its cytotoxicity to the B16-F10 melanoma cell lines, MTT reduction assay and morphological changes were observed. No cytotoxicity was found in horseradish methanol extract and SW 200. In conclusion, this research suggests the possibility of horseradish subcritical water may be useful as a natural whitening ingredient to be used in cosmetic products.

• Journal of Life Science
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• v.28 no.8
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• pp.908-916
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• 2018

Hair loses melanin with aging, which leads to hair graying. The change in hair color is caused by a reduction in tyrosinase activity and an accumulation of hydrogen peroxide ($H_2O_2$) in hair follicles. The purpose of this study was to investigate the effect of ethanolic extract of Oryza sativa (OREE) on melanin production and antioxidative activity in B16F1 cells. In this study, OREE showed low DPPH radical scavenging activity and reducing power. However, it displayed a strong antioxidative effect against intracellular $H_2O_2$ in live cells. OREE did not inhibit DOPA oxidation activity in vitro, but it increased tyrosinase activity at a concentration of $64{\mu}g/ml$. OREE at a concentration higher than $32{\mu}g/ml$ showed cell toxicity in B16F1 cells. However, OREE at a concentration higher than $8{\mu}g/ml$ not only increased melanin synthesis in a dose-dependent manner in B16F1 cells but also increased melanin synthesis in cells treated with $H_2O_2$ inhibiting melanin synthesis. To confirm the effect of OREE on melanin production, Western blot analysis was performed. The results revealed that OREE increased the expression levels of tyrosine hydroxylase and tyrosinase-related protein-2 (TRP-2) involved in melanin production in the $H_2O_2$-treated cells in which melanin production was inhibited. The findings suggest that OREE could improve melanin synthesis and be available for development of hair cosmetics aimed at improving melanin production.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.21 no.1
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• pp.70-82
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• 2008

Objective : The purpose of this study is to examine the effects of Baickbujasan(BB) on the skin damage and depigmentation. Method : The inhibition of tyrosinase activity, melanogenesis and cell viability in cultured B16 melanoma cells were measured. In order to test effects of reduction of melanogenesis, B16 F-10 mouse melanoma stem line was employed to extract melanin from cultured cell, where BB was added or not, and was dissolved in alkali for colorimetric analysis. Also, in order to test skin alteration in C57BL/6 after UV irradiation, the animals were grouped into a UV urradiation group and UV irradiation after BB application group. Dopa oxidase tissue staining was excuted to invesitage the change in the distribution of active melanin cell. The distribution of active melanin cell in inner skin of iNOS after damage from UVB irradiation and the manifestation condition of P53 which takes part in natural death of keratinocyte were examined. Result : The results indicate that BB has significant effects on tyrosinase activity, and melanogenesis in vivo test. BB seems to reduce C57BL/6, external dermatological damage, for instance, erythematous papule, eczema, loss of keratinocyte, reduction in pus, and relieves dermatological damages. Conclusion : BB can be applied externally for UV protection and depigmentation.

• Proceedings of the SCSK Conference
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• 2003.09b
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• pp.201-212
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• 2003

The heartwood extract of Artocarpus lakoocha Roxb., which contains a potent tyrosinase inhibitor oxyresveratrol, was evaluated for its melanin-reducing efficacy in both guinea pigs and human volunteers. After 4 week-daily application of the extract dissolved in propylene glycol to the back of guinea pigs, significant reduction in melanin content was detected, with the effect greater than 3% kojic acid and solvent propylene glycol (P < 0.05). The extract was subsequently tested in female volunteers (3 groups of 20 subjects) using a parallel clinical trial with self-control. The first group received the A. lakoocha solution in propylene glycol whereas the second and the third group respectively received 0.25% licorice extract and 3% kojic acid in the same solvent. The subject in each group twice daily applied the test solution on one arm whereas the remaining arm was applied with only propylene glycol (self-control) for 12 weeks. The A. lakoocha extract was found to be the most effective agent, giving the shortest onset of significant whitening after only 4 weeks of application (P < 0.05), followed by 3% kojic acid (8 weeks) and 0.25% licorice extract (10 weeks). The whitening effect also increased with time, with the highest extent observed with A. lakoocha at week 12. The in vitro antityrosinase activity of A. lakoocha extract decreased with time upon storage at room temperature but could be stabilized by a combination of several antioxidants. In conclusion, the heartwood extract of A. lakoocha appeared to have promising potential for use as an effective and economical skin-whitening agent.

• The Plant Pathology Journal
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• v.16 no.3
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• pp.125-129
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• 2000

A rapid and efficient assay to determine melanin biosynthesis inhibition of Magnaporthe grisea, a causal agent of the rice blast, by chemicals was developed. Wells in 24-well plates were loaded with spore suspension of the fungus and three known melanin biosynthesis inhibitors of KC10017, tricyclazole, and carpropamid. Subsequent color changes of mycelia and culture media in the wells were observed 7 days after incubation. The wells treated with KC10017 (an inhibitor of polyketide synthesis step and/or pentaketide cyclization step) became colorless, whereas tricyclazole (an inhibitor of 1, 3, 8-trihydroxynaphthalene reductase) or carpropamid (an inhibitor of scytalone dehydratase)-treated wells exhibited red color. They did not show any inhibitory effect on fungal growth. The inhibition of reaction steps prior to 1, 3, 6, 8-tetrahydroxynaphthalene formation was easily determined by colorless medium and mycelia. However, it was impossible to distinguish between inhibition of reduction steps and inhibition of dehydration steps by colors of the cultures. It was accomplished through HPLC analysis of the melanin biosynthesis-involving pentaketide metabolites accumulated by the inhibitors. Through screening of a number of synthetic chemicals using the in vitro assay, we could find a novel chemical group of melanin biosynthesis inhibitor.